Fig. 3. Mfn1 and Mfn2 protein expression in different cell lines, human and mouse tissues. (A) Extracts from COS-7 cells transiently transfected with various Mfn1 (GFP-Mfn1, Mfn1, antisense-Mfn1) and Mfn2- (Mfn2-myc and Mfn2^1-96-myc) expression constructs (as indicated) analyzed by western blotting using anti-Mfn1 or anti-myc antibodies. (B) Human Mfn1 protein detected as an 86 kDa band (arrow) in extracts from adult human heart, kidney (both from Clontech) and postnuclear supernatant from HeLa cell extracts probed with anti-Mfn1. Unspecific crossreacting band (asterisk). Same gel probed with anti-Tom40 antibodies as loading control (lower panel). (C) Mfn2 is abundantly expressed in human heart. Total cell extracts from yeast containing pGal1::Mfn2 plasmid and grown in either dextrose- or galactose-containing medium (left two lanes). Protein extracts from human kidney or heart (right two lanes) probed with Mfn2-specific polyclonal antibodies (two different exposures shown). Mfn2 runs as a doublet at around 86 kDa (arrow). Parallel loadings of same extracts probed with anti-Mfn1 antibodies, or anti-Tom40 and anti-actin as loading controls (lower three panels).